Method for introducing an ultraviolet light activated viral vector into the spinal column

a technology of ultraviolet light and activated viral vector, which is applied in the field of gene therapy, can solve the problems of reducing the range of motion and pain and/or disability, increasing the rate of second strand synthesis, and reducing the absorption of biomechanical shock, so as to facilitate the rebuilding of important components and effectively stimulate the transduction of uv activated viral vector.

Inactive Publication Date: 2010-04-27
UNIVERSITY OF ROCHESTER
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0016]A feature of preferred embodiments of the present invention the ability to overcome the problems involved with using traditional UV and γ-irradiation, by using locally administered UV, preferably long wavelength UV light, in order to induce the target cells to more effectively stimulate the transduction of a UV activated viral vector, such as recombinant adeno-associated virus r-AAV. In certain preferred embodiments, the desirable genetic material carried in the UV activated viral vector is then able to facilitate the rebuilding of important components of the FSU, such as GAG and collagen, or to effect bone formation resulting in fusion.

Problems solved by technology

This aging or degenerating, which is incompletely understood, generally results in decreased biomechanical shock absorption, increased range of motion and pain and / or disability.
The introduction of either the co-infection or the DNA damaging agents dramatically induces the rate limiting step of second strand synthesis, i.e. the second strand of DNA which is synthesized based on the vector inserted first strand.
However, making use of these DNA damaging agents is impractical because the administration of an adenovirus co-infection to a patient is not practical or desirable and the site specific and safety issues involved with using γ-irradiation are undesirable as well.
Unfortunately, no effective therapeutic method or apparatus was developed based on these experiments due to the long exposure times involved with using 254 nm UV radiation, the difficulties of delivering 254 nm UV radiation to a surgical target site, and the inability to position the 254 nm UV light source so as to allow effective penetration of a target cell.

Method used

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  • Method for introducing an ultraviolet light activated viral vector into the spinal column
  • Method for introducing an ultraviolet light activated viral vector into the spinal column
  • Method for introducing an ultraviolet light activated viral vector into the spinal column

Examples

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example 1

I. Methods

[0058]A. Isolation of Human Mesenchymal Stem Cells

[0059]Human Mesenchymal Stem Cells (hMSC) were isolated from patient blood samples harvested from the iliac crest. The blood samples were diluted in an equal volume of sterile Phosphate Buffered Saline (PBS). The diluted sample was then gently layered over 10 ml of Lymphoprep (Media Prep) in a 50 ml conical tube (Corning). The samples were then centrifuged at 1800 rpm for 30 minutes. This isolation protocol is a standard laboratory technique, and the resulting gradient that formed enabled the isolation of the hMSCs from the layer immediately above the Lymphoprep. The isolated fraction was placed into a new 50 ml conical tube, along with an additional 20 ml of sterile PBS. The sample was centrifuged at 1400 rpm for 8 minutes. The supernatant was removed the cell pellet was resuspended in 20 ml for fresh PBS, and centrifuged again for 8 minutes at 1400 rpm. Afterwards the supernatant was removed, the cell pellet was resuspend...

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Abstract

In accordance with the present invention, methods and structures are provided for the treatment of functional spinal unit injuries through the use of light activated gene therapy to induce bone fusion through the introduction of a desired gene into a patient's spinal tissue. Methods and structures are also provided for the utilization of ultraviolet light activated gene therapy to repair / rebuild an injured intervertebral disc through the introduction of a desired gene into a patient's spinal tissue. An implant system including a light probe and an implant with which r-AAV is integrated is also provided.

Description

RERFERENCE TO RELATED APPLICATIONS[0001]The present application claims the priority benefit under 35 U.S.C. §119(e) of Provisional Application No. 60 / 353,907, filed on Jan. 31, 2002. The present application is also related to Provisional Application No. 60 / 353,842, filed on Jan. 31, 2002, and U.S. application Ser. No. 10 / 357,271, filed on Jan. 31, 2003.GOVERNMENT INTEREST[0002]This invention was made with Government support under NIH Contract #AR45971, an RO1 grant awarded by NIAMS. The Government has certain rights in the invention.BACKGROUND OF THE INVENTION[0003]1. Field of the Invention[0004]The present invention relates to the field of gene therapy. According to the present invention, methods are provided for the treatment of functional spinal unit injuries through the use of light activated gene therapy to introduce a desired gene into a patient's tissue. An embodiment of the present invention includes methods for the utilization of light activated gene therapy to repair / rebui...

Claims

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Application Information

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Patent Type & Authority Patents(United States)
IPC IPC(8): A61N5/06A01N63/00A61K48/00C12N15/864
CPCA61K48/0008C12N15/86A61K48/0083A61K48/00C12N2750/14143
Inventor RUBERY, PAUL T.SCHWARZ, EDWARD M.
Owner UNIVERSITY OF ROCHESTER
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